Effect of siRNA against nerve growth factor (NGF) on growth and invasion of ovarian cancer cell

normal ovaries and in epithelial ovarian carcinomas. To examine the effects on ovarian cancer cell line (A2780) proliferation, invasion, and apoptosis after NGF down-regulation induced by lentivirus induced RNAi. Materials and Methods: The expression and localization of NGF, TrkA, p75, and VEGF in normal ovarian samples and in ovarian cancer samples were analyzed by RT-PCR and immunohistochemistry. NGF knockdown in A2780 was achieved by shRNA and cell proliferation, invasion, and apoptosis was examined by MTT, transwell assay, and flow cytometry, respectively. Results: Significantly higher levels of NGF, TrkA, and VEGF were observed in ovarian cancers versus normal ovary, while expression level of p75 maintained stable. In A2780 cells, NGF down-regulation significantly reduced the expression levels of TrkA and VEGF, but did not change that of p75. A significant decrease in proliferation and invasion, as well as an increase in apoptosis was also observed in NGF down-regulation cells. Conclusions: The expression of TrkA and VEGF is importantly induced by NGF. Knockdown of NGF promoted apoptosis, thus inhibited cellular proliferation and invasion in A2780 cells. Therefore NGF block may be a potential therapeutic strategy to treat ovarian cancer.

NGF; NGF siRNA; TrkA; p57; VEGF; Ovarian cancer.

[1] Urzúa U., Tapia V., Geraldo M.P., Selman A., Vega M., Romero C.: “Nerve growth factor stimulates cellular proliferation of human epithelial ovarian cancer”. Horm. Metab. Res., 2012, 44, 656.

[2] Bast R., Hennessy B., Mills G.B.: “The biology of ovarian cancer: new opportunities for translation”. Nat. Rev. Cancer, 2009, 9, 415.

[3] Reynolds L., Grazul-Bilska A., Redmer D.: “Angiogenesis in the female reproductive organs: pathological implications”. Int. J. Exp- Pathol., 2002, 83, 151.

[4] Julio-Pieper M., Lara H., Bravo J., Romero C.: “Effects of nerve growth factor (NGF) on blood vessels area and expression of the angiogenic factors VEGF and TGF beta1 in the rat ovary”. Reprod. Biol. Endocrinol., 2006, 4, 57.

[5] Levi-Montalcini R.: “The nerve growth factor: its mode of action on sensory and sympathetic nerve cells”. Harvey Lect., 1966, 60, 217.

[6] Oelmann E., Srete L., Schuller I., Serve H., Koenigsmann M., Wiedenmann B., et al.: “Nerve growth factor stimulates clonal growth of human lung cancer cell lines and a human glioblastoma cell line expressing high-affinity nerve growth factor binding sites involving tyrosine kinase signaling”. Cancer Res., 1995, 55, 2212.

[7] Sortino M.A., Condorelli F., Vancheri C., Chiarenza A., Bernardini R., Consoli U., et al.: “Mitogenic effect of nerve growth factor(NGF) in LNCaP prostate adenocarcinoma cells: role of the high and low affinity NGF receptors”. Mol. Endocrinol., 2000, 14, 124.

[8] Tagliabue E., Castiglioni F., Ghirelli C., Modugno M., Asnaghi L., Somenzi G., et al.: “Nerve growth factor cooperates with p185 (HER2) in activating growth of human breast carcinoma cells”. J. Biol. Chem., 2000, 275, 5388.

[9] Dissen G., Hirshfield A., Malamed S., Ojeda S.: “Expression of neurotrophins and their receptors in the mammalian ovary is developmentally regulated: changes at the time of folliculogenesis”. Endocrinology, 1995, 136, 4681.

[10] Dissen G., Romero C., Hirshfield A., Ojeda S.R.: “Nerve growth factor is required for early follicular development in the mammalian ovary”. Endocrinology, 2001, 142, 2078.

[11] Cantarella G., Lempereur L., Presta M., Ribatti D., Lombardo G., Lazarovici P., et al.: “Nerve growth factor-endothelial cell interaction leads to angiogenesis in vitro and in vivo”. FASEB J., 2002, 16, 1307.

[12] Tapia V., Gabler F., Munoz M., Yaziqi R., Paredes A., Selman A., et al.: “Tyrosine kinase A receptor (trkA): a potencial marker in epithelial ovarian cancer”. Gynecol. Oncol., 2011, 121, 13.

[13] Calza L., Giardino L., Giuliani A., Aloe L., Levi-Montalcini R.: “Nerve growth factor control of neuronal expression of angiogenetic and vasoactive factors”. Proc. Natl. Acad. Sci. USA, 2001, 98, 4160.

[14] Vera C., Tapia V., Vega M., Romero C.: “Role of nerve growth factor and its TRKA receptor in normal ovarian and epithelial ovarian cancer angiogenesis”. J. Ovarian Res., 2014, 7, 82.

[15] Kaplan D., Miller F.D.: “Signal transduction by the neurotrophin receptors”. Curr. Opin. Cell. Biol., 1997, 9, 213.

[16] Campos X., Muñoz Y., Selman A., Yazigi R., Moyano L., Weinstein-Oppenheimer C., et al.: “Nerve growth factor and its high-affinity receptor TrkA participate in the control of vascular endothelial growth factor expression in epithelial ovarian cancer”. Gynecol. Oncol., 2007, 104, 168.

[17] Zhang X., LaRussa V., Reiser J.: “Transduction of bone-marrowderived mesenchymal stem cells by using lentivirus vectors pseudotypedwith modified RD114 envelope glycoproteins”. J.Virol., 2004, 78, 1219.

[18] Gómez-Raposo C., Mendiola M., Barriuso J., Casado E., Hardisson D., Redondo A.: “Angiogenesis and ovarian cancer”. Clin. Transl. Oncol., 2009, 11, 564.